Up to 100 million Dengue virus infections occur annually world wide. Associated pathologies (Dengue Hemorrhagic Fever, Dengue Shock Syndrome) result in >20,000 deaths annually, primarily in children. Dengue virus is endemic to tropical/subtropical regions, with high incidence rates in developing countries. Currently, no specific treatments or vaccines are available. Moreover, rapid, sensitive and specific diagnostics are lacking. Antibody ELISA tests lack specificity and require a host immune response; tissue culture assays require specialized laboratories and long incubation times. Phase I of this SBIR demonstrated the feasibility of linking a simple nucleic acid isolation technology to an isothermal method of nucleic acid amplification and a liposome-microchip biosensor detection technology for the clinical detection/subtyping of Dengue virus. This technology linkage permits specific and sensitive detection of Dengue infection in patients within 5 hours, has minimal equipment requirements, and can be provided at costs compatible with endemic area economies. Phase II proposes continued development/optimization of this assay system. Phase I design specifications will be used to produce and validate a prototype assay, produced in Jab batch quantities. Identified design and production modifications will be implemented to produce pilot batch lots of the assay that will be validated and clinically evaluated. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE